Hubbuch Jürgen, Kula Maria Regina
Institut für Biotechnologie 2, Forschungszentrum Jülich, 52425 Jülich, Germany.
Bioprocess Biosyst Eng. 2008 Apr;31(3):241-59. doi: 10.1007/s00449-008-0197-5. Epub 2008 Jan 15.
In recent years, confocal laser scanning microscopy has been developed into a non-invasive tool to probe intra-particle profiles of protein in chromatographic adsorbents. A necessary prerequisite when using this technique lies in the labeling of proteins with fluorescent probes. The quality of the obtained results is thus strongly dependent on the probes used, its sensitivity on experimental parameters and the change of protein characteristics upon binding. In this review, the fundamental issues when using fluorescent probes are described, before giving a critical evaluation on published literature in the field of confocal laser scanning microscopy for the analysis of chromatographic principles.
近年来,共聚焦激光扫描显微镜已发展成为一种用于探测色谱吸附剂中蛋白质颗粒内分布的非侵入性工具。使用该技术的一个必要前提是用荧光探针标记蛋白质。因此,所获得结果的质量在很大程度上取决于所使用的探针、其对实验参数的敏感性以及结合后蛋白质特性的变化。在本综述中,先描述了使用荧光探针时的基本问题,然后对共聚焦激光扫描显微镜用于分析色谱原理领域的已发表文献进行了批判性评价。